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PHYSIOLOGY  OF  THE  PHENOLS 


RECAP 


BT 

HARRY  DUBIN 


THESIS  PRESENTED  TO  THE  FACULTY  OF  THE   GRADUATE 
SCHOOL  OF  THE  UNIVERSITY  OF  PENNSYLVANIA,  IN 
PARTIAL  FULFILMENT    OF  THE    REQUIRE- 
MENTS FOR  THE  DEGREE  OF  DOC- 
TOR OF  PHILOSOPHY 


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PHYSIOLOGY   OF   THE   PHENOLS 


BY 

HARRY  DUBIN 


THESIS  PRESENTED  TO  THE  FACULTY  OF  THE  GRADUATE 
SCHOOL  OF  THE  UNIVERSITY  OF  PENNSYLVANIA,  IN 
PARTIAL  FULFILMENT  OF  THE  REQUIRE- 
MENTS FOR  THE  DEGREE  OF  DOC- 
TOR OF  PHILOSOPHY 


Reprinted  from  The  Journal  of  Biological  Chemistry,  Vol.  XXVI,  No.  1,  1916 


PHYSIOLOGY  OF  THE  PHENOLS.* 

By  harry  DUBIN. 

{From  the  Department  of  Physiological  Chemistry  and  the  John  Herr  Musser 

Department  of  Research  Medicine,  University  of 

Pennsylvania,  Philadelphia. ) 

(Received  for  publication,  June  1,  1916.) 
INTRODUCTION. 

The  work  of  Baumann,  Brieger,  Salkowski,  and  others  has 
seemingly  solved  the  problem  of  the  origin  of  phenols  in  urine. 
It  is  clear  that  the  urinary  phenols  arise  from  intestinal  putrefac- 
tion, and  that  they  .are  derived  from  the  tyrosine  portion  of  the 
protein  molecule.  However,  with  the  exception  of  the  work  of 
Folin,^  little  has  been  done — due  probably  to  the  lack  of  a  suit- 
able method — to  determine  (1)  the  extent  to  which  phenols  may 
be  formed  under  both  normal  and  pathological  conditions,  and 
(2)  the  relationship  between  the  free  and  conjugated  phenols.  It 
was  with  these  objects  in  view  that  the  present  work  was  under- 
taken. 

HISTORICAL. 

The  literature  has  to  a  certain  extent  been  reviewed  by  Folin/  so  that 
it  will  be  necessary  only  to  record  the  results  of  some  recent  investigations. 

The  elimination  of  phenols  is  increased  by  absorption  from  wounds  and 
abscesses. 2 

The  relative  ability  of  the  various  organs  to  dispose  of  phenol  is  as 
follows:  liver,  kidney,  muscle,  brain,  and  blood;  i.e.,  the  liver  has  the 
greatest  capacity  for  conjugating  phenols,  while  the  blood  has  the  least. 
Only  the  epithelia  of  the  intestinal  tract  give  results  comparable  to  those 
obtained  with  the  liver. ^  The  conjugating  function  of  the  liver  is  not  much 
impaired  by  disease.^ 

*  Dissertation  for  the  degree  of  Ph.D.,  University  of  Pennsylvania, 
1916. 

iFolin,  O.,  and  Denis,  W.,  /.  Biol.  Chem.,  1915,  xxii,  309. 

2  Hammarsten,  O.,  Physiological  Chemistry,  New  York,   1911,  689. 

3  Herter,  C.  A.,  and  Wakeman,  A.  J.,  J.  Exp.  Med.,  1899,  iv,  307. 

69 


70  Physiology  of  Phenols 

On  starving,  the  phenols,  after  an  initial  fall,  increase  considerably.^ 
Muller^  reported  the  case  of  a  man  who  showed  an  increase  of  155  mg.  of 
phenol  on  the  9th  day  of  starvation.  This  was  held  to  be  due  to  putre- 
faction of  intestinal  secretions.  If,  while  an  animal  is  starving,  phlor- 
hizin  be  given,  there  is  an  increase  in  the  output  of  phenols,  but  not  until 
sugar  makes  its  appearance.^  It  is  the  increased  protein  breakdown, 
rather  than  intestinal  putrefaction,  that  accounts,  under  these  circum- 
stances, for  the  increase  in  the  elimination  of  phenols.  Lewin^  observed 
in  rabbits  an  increase  from  6.91  mg.  of  phenol  to  16.35  mg.  of  phenol  per 
day,  on  giving  0.8  gm.  of  phlorhizin  subcutaneously.  In  man  there  was 
an  increase  of  two  or  three  times  the  original  amount  of  phenol  upon 
administering  0.25  gm.  of  phlorhizin  per  os  or  subcutaneously. 

It  is  well  known  that  if  a  measured  amount  of  phenol  be  ingested  it 
cannot  be  recovered  quantitatively  in  the  urine,  and  it  is  believed  that 
the  missing  fraction  is  burned.  Tauber^  fed  phenol  to  a  dog  and  found 
that  as  the  dose  was  decreased,  the  amount  of  phenol  oxidized  increased. 
Thus,  he  found  no  phenol  in  the  fore-period,  either  in  the  urine  or  in  the 
feces.  On  feeding  0.24  gm.  of  phenol  in  water  per  os,  he  found  110  mg.  of 
phenol  in  the  urine,  and  only  9  mg.  in  the  feces.  In  other  words,  about 
63  per  cent  of  the  ingested  phenol  was  oxidized  in  the  body  through  oxalic 
acid  to  carbon  dioxide.  Heffter^  states  that  after  feeding  phenol,  there  is 
an  increase  in  phenolsulfuric  acid  but  not  in  free  phenol.  On  the  other 
hand,  Reale^  reports  that  poisoning  with  large  amounts  of  phenol  results 
in  the  presence  of  free,  as  well  as  combined,  phenol  in  the  urine. 

Jonescu^"  found  that  after  feeding  p-cresol  to  dogs,  kept  on  a  diet  of 
horse  meat,  only  about  25  per  cent  of  the  amount  ingested  is  eliminated, 
and  that  the  elimination  is  complete  in  24  hours.  Taking  an  average  of 
nine  persons,  Siegfried  and  Zimmermanii  showed  that  of  the  total  phenols 
eliminated,  58.1  per  cent  is  p-cresol,  and  41.9  per  cent  is  phenol.  They^^ 
found  also  that  jo-cresol,  fed  to  dogs,  is  largely  changed  to  phenol.  Thus, 
on  feeding  0.5  gm.  of  p-cresol  with  0.5  gm.  of  sodium  bicarbonate  on  4 
successive  days,  32  per  cent  of  the  total  amount  fed  was  recovered  in  the 
ufine.     Similarly,  on  feeding  0.8  gm.  of  p-cresol,  48  per  cent  was  recovered. 

The  occurrence  of  the  phenols  in  the  urine  upon  disinfection  of  the 
intestine  is  shown  in  an  experiment  carried  out  by  Baumann.^^    A  dog 

*  Herter,  C.  A.,  Chemical  Pathology,  Philadelphia,  1902,  425. 
^Miiller,  F.,  Berl.  Idin.  Woch.,  1887,  xxiv,  405,  436. 
«  Lewin,  C,  Beitr.  chem.  Phys.  u.  Path.,  1902,  i,  472. 

7  Tauber,  E.,  Z.  physiol.  Chem.,  1878-79,  ii,  366. 

8  Heffter,  A.,  Ergebn.  Physiol.,  1905,  iv,  242. 

"  Reale,  E.,  Abstr.,  Centr.  klin.  Med.,  1891,  xii,  487,  quoted  iromJahresber. 
Tierchem.,  1891,  xxi,  401  (orig.,  Gaz.  clin.,  1890,  i,  2). 
10  Jonescu,  D.,  Biochem.  Z.,  1906,  i,  399. 

^1  Siegfried,  M.,  and  Zimmerman,  R.,  Biochem.  Z.,  1911,  xxxiv,  471. 
'-  Siegfried  and  Zimmerman,  Biochem.  Z.,   1912,  xlvi,  210. 
12  Baumann,  E.,  Z.  physiol.  Chem.,  1886,  x,  123. 


Harry  Dubin  71 

receiving  only  water  for  2  days  was  given  2  gm.  of  calomel  on  the  2nd 
day.  As  on  the  4th  day  ethereal  sulfates  were  still  present  in  the  urine, 
2  gm.  of  caiomel  were  again  given.  The  urine  of  the  2  following  days  was 
free  of  ethereal  sulfates,  and  gave  no  test  for  phenol  or  indole.  On  the 
6th  day,  the  dog  received  5  gm.  of  tyrosine,  but  no  formation  of  ethereal 
sulfates  resulted;  neither  was  there  an  increase  of  hydroxy  acids. 

Wohlgemuth^*  fed  8  gm.  of  tyrosine  to  a  rabbit  and  was  able  to  recover 
a  little  less  than  2  gm.  of  it.  Dakin^^  reports  no  increase  of  phenolic  sub- 
stances after  giving  tyrosine  or  phenylalanine.  After  feeding  tyrosine  to 
cats,  he  recovered  by  crystallization  from  the  urine  very  small  amounts 
of  tyrosine.  He  found  also  that  with  smaller  doses  no  unchanged  tyrosine 
could  be  recovered  in  the  urine.  Brieger^^  could  find  no  tyrosine  in  the 
feces  or  urine  after  giving  20  gm.  of  tyrosine  to  a  man  weighing  about  50 
kilos.  He  did,  however,  find  an  increase  in  the  phenols,  and  although  the 
patient  was  constipated  for  2  days  after  taking  the  tyrosine,  Brieger  held 
that  the  increase  in  phenols  could  not  be  due  to  constipation  alone. 
Brieger's  figures  follow: 


Day. 
1 

Pheno]. 
gm. 

.  0.0159 

2 

,  0.0225 

3  ... 

.  0.0223 

4 

,  0.0182 

20  gm. 
5 

of  tyrosine 

ingested  in 

two 

portions. 
,  0.0493 

6 

.  0.1576 

7.... 

.  0.0851 

8.... 

.  0.0609 

9.... 

.  0.0348 

Results  obtained  with  animals  and  with  man  have  led  Folin^  to  con- 
clude that  the  excretion  of  total  phenol  products  in  the  urine  appears  to 
be  much  greater  than  is  indicated  by  the  phenol  figures  previously  re- 
corded in  the  literature.  He  shows  further  that  the  phenols  are  not 
quantitatively  converted  into  conjugated  phenols,  so  that  the  detoxica- 
tion  process  involved  in  such  conjugations  appears  to  furnish  only  a  partial 
protection  against  the  toxic  effect  of  the  phenol  products  formed  by  putre- 
faction in  the  intestinal  tract.  Finally,  the  total  phenol  excretion  tends 
to  vary  directly,  but  not  proportionally,  with  the  protein  intake. 

Some  of  the  work  described  in  this  summary  has  been  confirmed  by  our 
findings,  while  some  has  not.     The  value  of  our  figures  lies  in  the  fact  that 


"  Wohlgemuth,  J.,  Ber.  chem.  Ges.,  1905,  xxxviii,  2064. 

15  Dakin,  H.  D.,  J.  Biol.  Chem.,  1910-11,  viii,  28. 

"Brieger,  L.,  Z.  physiol.  Chem.,  1878-79,  ii,  241;  1879,  ill,  134. 


72  Physiology  of  Phenols  ' 

they  have  been  obtained  by  a  more  accurate  method,  that  of  Folin/^  which 
permitted  not  only  a  definite  quantitative  determination,  but  also  the 
study  of  the  relationship  between  the  free  and  the  conjugated  phenols. 

Methods. 

For  phenols,  free  and  conjugated,  the  technique  of  Fohn^^  was 
used,  bearing  in  mind  the  following  precautions:  (a)  In  determin- 
ing total  phenols,  a  few  glass  beads  should  be  used  to  prevent  the 
liquid  in  the  test-tube  from  over-boiling,  during  the  heating  just 
prior  to  placing  the  tube  in  the  boiling  water  bath,  (b)  The  phos- 
photungstic  phosphomolybdic  acid  reagent  is  prepared  by  boiling 
the  various  chemicals  with  750  cc.  of  water  instead  of  75  cc,  the 
amount  stated  in  Folin's  description,  which  was  an  error,  (c)  The 
stock  phenol  solution  from  which  the  standard  is  prepared 
holds  its  strength  for  a  long  period  of  time,  whereas  the  standard 
phenol  solution  itself  deteriorates,  and  should  therefore  be  pre- 
pared freshly  every  4  or  5  days,  (d)  The  color  obtained  with  the 
standard  solution  is  not  absolutely  blue,  but  has  a  greenish  tinge. 
This  defect  is  remedied  by  filtration,  without  in  any  way  affecting 
the  colorimeter  reading.  Therefore,  after  standing  the  required 
20  minutes,  the  standard  is  filtered  through  inexpensive  coarse 
jfilter  paper,^^  and  the  colorimeter  readings  are  made  at  once. 

With  these  precautions  we  have  found  the  method  accurate, 
rapid,  and  easy  of  application. 

EXPERIMENTAL. 

In  this  investigation  an  effort  has  been  made  to  determine  the 
behavior  of  the  animal  body,  with  respect  to  the  formation  and 
elimination  of  phenols,  under  both  normal  and  pathological  con- 
ditions. Dogs  were  chosen  as  being  best  suited  for  the  work. 
After  a  period  of  normal  observation,  the  animals  were  operated 
upon,!^  and  the  desired  pathological  conditions  produced.  All 
operations  were  done  under  ether  anesthesia.  In  all,  eight  opera- 
tions were  performed,  two  each  of:  (1)  Eck  fistula;  {2)  intestinal 
obstruction;  (S)  exclusion  of  bile  from  the  intestinal  tract;  (4)  exclu- 

1'  Folin  and  Denis,  ./.  Biol.  Chem.,  1915,  xxii,  305. 
18  Arthur  H.  Thomas  Co.,  No.  27756,  125  mm. 

1' For  the  operative  work,  I  am  indebted  to  Dr.  Max  Minor  Peet,  of 
the  Department  of  Surgical  Research. 


Harry  Dubin  73 

sion  of  pancreatic  juice  from  the  intestinal  tract.     The  technique 
employed  in  each  operation  was  as  follows: 

1.  Eck  Fistula. — An  artificial  anastomosis  between  the  portal 
vein  and  the  inferior  vena  cava,  with  ligation  of  the  portal  vein 
at  the  hilus  of  the  liver,  was  made  according  to  the  technique  de- 
scribed by  Peet.2''' 

S.  Intestinal  Obstruction. — Almost  complete  obstruction  of  the 
ileum  was  obtained  about  6  inches  from  the  ileocecal  valve  by 
detaching  from  the  external  sheath  of  the  rectus  a  strip  about  | 
inch  wide  and  1  inch  long,  passing  it  through  the  mesenteric  at- 
tachment around  the  gut,  overlapping,  and  suturing  the  two  ends 
of  the  fascia  firmly  together. 

3.  Exclusion  of  Bile  from  the  Intestinal  Tract. — The  common  bile 
duct  was  tied  in  three  places,  cut  between  the  ligatures,  and  part 
of  the  omentum  sewed  in  between  the  cut  ends  to  prevent  a  pos- 
sible reunion. 

4-  Exclusion  of  Pancreatic  Juice  from  the  Intestinal  Tract. — In 
one  animal  two  ducts  were  found,  while  in  another,  three  ducts 
were  present.  Each  duct  was  divided  between  ligatures  and  part 
of  the  omentum  interposed  between  the  cut  ends,  for  the  reason 
just  mentione4- 

Plan  of  Investigation. 

The  general  plan  of  the  investigation  was  to  study  the  daily  out- 
put of  urinary  and  fecal  phenols  in  dogs  under  the  conditions  out-  . 
lined.  The  elimination  of  phenols,  under  the  influence  of  fasting, 
catharsis,  and  phlorhizin,  was  also  studied.  Inasmuch  as  only 
minimal  amounts  of  phenol  were  found  in  the  feces — amounts 
that  could  have  no  bearing  on  the  iinal  results — these  determina- 
tions were  dispensed  with. 

The  dogs,  kept  in  metabolism  cages,  were  fed  on  a  standard  diet, 
calorifically  sufficient,  and  containing  about  1  gm.  of  nitrogen  per 
kilo  so  as  to  make  certain  of  an  excess  of  food  in  the  intestine. 
This  diet  consisted  of  meat,  lard,  bread  crumbs,  sugar,  salt,  and 
sufficient  bone  ash  to  insure  a  well  formed  stool.  The  food  was 
mixed  with  400  cc.  of  water,  and  enough  additional  water  was 
given  separately  to  bring  the  total  daily  intake  up  to  about  600 
cc.     The  animals  were  placed  on  this  diet  3  or  4  days  before  the 

2«  Peet,  M.  M.,  Ann.  Surg.,  1914,  Ix,  601. 


74 


Physiology  of  Phenols 


beginning  of  an  experiment.  Female  dogs  only  were  used,  the 
urine  being  collected  by  catheter  every  morning  at  the  same  hour 
and  diluted  to  1,000  cc.  For  a  time  the  volume  of  urine  excreted 
was  noted,  but  this  was  later  omitted  for  the  reason  that  no  par- 
ticular relationship  was  seen  between  the  volume  of  urine  and  the 
amount  of  phenols  eliminated,  except  as  noted  in  Table  I.  The 
body  weight  was  recorded  daily  immediately  after  catheteriza- 
tion. Tyrosine  was  administered  per  os,  either  suspended  in 
water  or  mixed  with  the  food.  Phenol  (Merck  reagent)  and  p-cresol 
(Kahlbaum)  were  given  per  os  in  water. 


TABLE  I. 

Eck  Fistula.     Dog  15-58. 

Influence  of  Water  and  Eck  Fistula  on  Phenol  Excretion. 


1 

'o 

S.2 

:3  f-i 

Total 

N. 

Phenols. 

Weight. 

Date. 

r  73 

Remarks. 

> 

Free. 

Total. 

6 

-3   o 
O   '^ 

o 

1915 

cc. 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Oct.  19 

600 

8.9 

0.158 

0.181 

87 

13 

12.4 

* 

"     20 

910 

9.7 

0.165 

0.192 

86 

14 

12.4 

"      21 

695 

6.1 

0.163 

0.188 

87 

13 

12.5 

"      22 

910 

9.7 

0.165 

0.190 

87 

13 

12.7 

"     23 

745 

10.4 

0.163 

0.188 

87 

13 

12.6 

"     24 

550 

9.9 

0.161 

0.186 

87 

13 

12.5 

"     25 

415 

9.9 

0.173 

0.203 

85 

15 

12.7 

Water  intake  reduced  to 
300  cc. 

"     26 

405 

9.8 

0.177 

0.205 

86 

14 

12.7 

"     27 

345 

9.9 

0.173 

0.203 

85 

15 

12.8 

"     28 

450 

9.9 

0.174 

0.203 

86 

14 

12.9 

"     29 

450 

9.9 

0.172 

0.201 

86 

14 

12.9 

"     30 

440 

9.8 

0.172 

0.203 

85 

15 

13.0 

"     31 

455 

9.9 

0.170 

0.200 

85 

15 

13.0 

Nov.  10 

Eck  fistula. 

"      11 

400 

10.2 

0.187 

0.203 

92 

8 

12.7 

Post-operative  condi- 
tion good. 

"      12 

250 

9.9 

0.186 

0.204 

92 

8 

12.6 

"      13 

350 

10.3 

0.188 

0.204 

92 

8 

12.9 

"     14 

360 

10.6 

0.197 

0.209 

94 

6 

13.1 

"      15 

590 

10.7 

0.205 

0.213 

96 

4 

13.2 

"      16 

570 

10,6 

0.199 

0.211 

94 

6 

13.1 

u     17 

480 

10.6 

0.200 

0.213 

94 

6 

13.0 

Harry  Dubin  75 

Eck  Fistula. 

Experiment  A-1. — Dog  15-58  (Table  I).  This  experiment 
showed  several  interesting  points.  The  output  of  phenols  from 
day  to  day  was  quite  constant,  as  was  also  the  case  in  all  of  our 
subsequent  work.  Withdrawing  water  caused  a  drop  in  the  vol- 
ume of  urine  with  a  consequent  rise  in  the  output  of  phenols. 
However,  it  was  seen  that  where  the  urine  varied  normally  from 
day  to  day,  there  was  little  or  no  effect  upon  the  phenols.  After 
Eck  fistula,  the  free  phenols  represented  from  92  to  96  per  cent 
of  the  total,  the  amount  of  the  latter  being  practically  unchanged. 
On  Nov.  17,  because  of  the  presence  of  blood  in  the  urine,  the 
dog  was  placed  on  a  kennel  diet. 

Experiment  A-2. — Dog  15-58  (Table  II).  2  weeks  later,  on 
Dec.  1,  the  regular  diet  was  resumed,  the  experiment  being  started 
Dec.  6.  For  some  reason,  which  we  have  been  unable  to  explain 
— unless  it  is  that  the  food  is  digested  more  rapidly — the  absolute 
amount  of  total  phenols  was  decreased  in  the  second  period,  but 
the  free  phenols  still  represented  from  96  to  98  per  cent  of  the 
total.  1  gm.  of  phenol  was  fed  per  os  with  the  result  that  68.7 
per  cent  of  the  amount  ingested  was  eliminated.  Both  free  and 
conjugated  phenols  were  increased.  The  free  phenols  represented 
only  40  per  cent  of  the  total,  indicating  that  the  large  dose  of 
phenol  had  called  forth,  to  an  increased  extent,  the  protective 
mechanism  of  the  body.  A  second  feeding  gave  similar  results. 
Feeding  1  gm.  of  p-cresol  resulted  in  the  elimination  of  50.6  per 
cent  of  the  amount  given.  Here  also  both  free  and  conjugated 
phenols  were  increased.  The  free  phenols  represented  only  31 
per  cent  of  the  total.  Repetition  of  this  feeding  gave  somewhat 
similar  results.  In  this  experiment  and  in  all  subsequent  ones,  it 
was  noted  that  the  phenol  and  p-cresol  administered  were  elimi- 
nated entirely  within  24  hours. 

The  results  obtained  on  feeding  tyrosine  have  a  peculiar  inter- 
est. For  example.  Dog  15-58  (Table  II),  receiving  400  gm.  of 
meat — equivalent  to  3.20  gm.  of  tyrosine,^!  or  1.65  gm.  of  phenol 
— eliminated  daily  a  total  of  only  0.159  gm.  of  phenol.  How- 
ever, on  feeding  5  gm.  of  tyrosine — equivalent  to  2.57  gm.  of 
phenol — there  was  a  rise  in  both  free  and  total  phenols,  while  the 

21  Folin  and  Denis,  /.  Biol.  Chem.,  1912,  xii,  246. 


76 


Physiology  of  Phenols 


TABLE  II. 

Eck  Fistula.     Dog  15-58. 
Influence  of  Phenol,  p-Cresol,  and  Tyrosine  on  Phenol  Excretion  after  Eck 

Fistula. 


Total 

N. 

Phenols. 

Weight. 

Date. 

1  TS 

Remarks. 

Free. 

Total. 

6 

u 

1915 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Dec.    6 

9.0 

0.143 

0.147 

97 

3 

13.9 

Cage  water  resumed. 

7 

9.0 

0.147 

0.152 

97 

3 

13.9 

8 

9.1 

0.334 

0.836 

(0.787)* 
(78.7%) 

40 

60 

14.0 

1.000  gm.  phenol  given 
in  water  per  os. 

9 

9.1 

0.152 

0.156 

97 

3 

"      10 

9.1 

0.153 

0.157 

97 

3 

14.2 

"      11 

9.2 

0.222 

0.517 
(0.360) 

(72.0%) 

41 

59 

14.2 

0.500  gm.  phenol  given 
in  water  per  os. 

"      12 

9.3 

0.154 

0.159 

97 

3 

14.2 

"      13 

9.5 

0.156 

0.159 

98 

2 

14.4 

"      14 

9.5 

0.188 

0.600 
(0.441) 

(50.6%) 

32 

68 

14.5 

1.000  gm.  p-cresol  (0.870 
gm.  phenol)  given  in 
water  per  os. 

"      15 

9.4 

0.154 

0.159 

97 

3 

14.5 

"      16 

9.5 

0.174 

0.541 
(0.381) 

(43.8%) 

34 

66 

14.5 

p-Cresol  fed  as  on  Dec. 
14. 

"      17 

9.7 

0.154 

0.157 

98 

2 

14.5 

"      18 

10.3 

0.420 

0.603 

(0.446) 

(17.7%) 

70 

30 

14.7 

5.000  gm.  tyrosine  (2.570 
gm.  phenol)  given  in 
water  per  os. 

"      19 

9.6 

0.152 

0.156 

97 

3 

14.7 

*  The  figures  in  the  first  parenthesis  represent  "extra"  phenols  elimi- 
nated; those  in  the  second  parenthesis  denote  the  output  of  "extra"  phenols 
in  terms  of  percentage  of  the  amount  ingested. 


"extra"  phenol  eliminated  was  only  0.446  gm.,  or  17.7  per  cent,  of 
the  amount  ingested.  In  other  words — and  it  was  to  be  expected — 
tyrosine,  as  present  in  meat,  did  not  give  rise  to  as  much  phenol 
as  did  free  tyrosine.  Similarly,  phenol,  administered  as  tyro- 
sine or  p-cresol,  did  not  give  rise  to  as  much  phenol  as  did  native 


Harry  Dubin 


77 


'phenol.  It  was  observed  that  tyrosine,  like  phenol  and  p-cresol, 
is  eliminated"  entirely  within  24  hours.  A  greater  conjugation 
was  also  seen,  though  not  as  large  as  that  produced  by  phenol  and 
p-cresol.  A  slight  rise  in  the  nitrogen  elimination  was  also  noted. 
At  no  time  was  it  possible  to  demonstrate  unchanged  tyrosine, 
either  in  the  urine  or  the  feces. 

Experiment  AS. — Dog   15-58   (Table  III).     1  gm.  of  phlor- 
hizin  rubbed  up  in  10  cc.  of  olive  oil  was  injected  subcutaneously 

TABLE  III. 

Eck  Fistula.     Dog  15-58. 

Influence  of  Phlorhizin,  Fasting,  and  Catharsis  on  Phenol  Excretion  after 


Eck  Fistula. 

Total 

N. 

Phenols. 

Weight. 

Date. 

3-d 

Remarks. 

Free. 

Total. 

u 
fo 

1915 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Dec.  19 

9.6 

0.152 

0.156 

97 

3 

14.7 

"      20 

13.1 

0.203 

0.277 

73 

27 

14.8 

1  gm.  phlorhizin. 

"      21 

15.5 

0,217 

0.281 

77 

23 

14.7 

-1        a                 a 

"      22 

13.9. 

0.171 

0.210 

82 

18 

14.4 

Sugar  in  urine  Dec.  20-23, 
inckisive. 

"      23 

9.9 

0.154 

0.159 

97 

3 

14.3 

Animal  placed  on  kennel 
diet  till  Dec.  31,  when 
regular  diet  was  resumed. 

1916 

' 

Jan.    5 

10.2 

0.150 

0.158 

95 

5 

14.7 

"        6 

5.2 

0.077 

0.111 

70 

30 

14.9 

Animal  fasting;  500  cc.  of 
water  given  in  cage. 

7 

4.8 

0.068 

0.091 

74 

26 

14.4 

■      "        8 

4.4 

0.068 

0.092 

74 

26 

13.9 

9 

4.0 

0.068 

0.091 

75 

25 

13.6 

"      10 

3.6 

0.067 

0.090 

74 

26 

13.4 

"      11 

3.5 

0.066 

0.091 

73 

27 

13.2 

"      12 

3  gm.  calomel  given;  cage 
urine  contaminated;  blad- 
der urine  contained  phe- 
nols. 

"      13 

3.4 

0.066 

0.089 

74 

26 

12.9 

"      14 

3.3 

0.066 

0  091 

73 

27 

12.7 

3  gm.  calomel  given. 

"      25 

3.0 

0  065 

0.089 

74 

I 

26 

11.5 

Animal  still  fasting. 

78 


Physiology  of  Phenols 


on  2  successive  days.  The  result  was  an  increase  in  both  total 
and  free  phenols,  the  latter  representing  73  per  cent  of  the  total. 
Sugar  appeared  in  the  urine,  just  as  it  did  later,  under  similar  cir- 

TABLE  IV. 

Eck  Fistula.     Dog  16-56. 

Influence  of  Phenol,  p-Cresol,  and  Tyrosine  on  Phenol  Excretion  before  and 

after  Eck  Fistula. 


Total 

N. 

Phenols.     , 

Weight. 

Date. 

a'6 

Remarks. 

Free. 

Total. 

CD 

1916 

gm. 

<;?«. 

gm. 

per 
cent 

per 
cent 

kg. 

May    9 

10.3 

0.176 

0.235 

75 

25 

12.3 

"      10 

1 

9.6 

0.385 

0.782 

(0.547) 

(54.7%) 

49 

51 

12.4 

1.000  gm.  phenol  given 
in  water  per  os. 

"      11 

10.1 

0.179 

0.241 

74 

26 

12.4 

"       12 

10.3 

0.435 

0.606 
(0.365) 

(14.2%) 

72 

28 

12.5 

5.000  gm.  tyrosine  (2.57 
gm.  phenol)  mixed 
with  food. 

"      13 

10.2 

0.176 

0.233 

75 

25 

12.6 

"      14 

10.2 

0.210 

0.556 
(0  323) 

(37.1%) 

38 

62 

12.5 

1.000  gm.  p-cresol  (0.870 
gm.  phenol)  given  in 
water  per  os. 

"      15 

Eck  fistula;  post-oper- 
ative condition  good. 

"      16 

10.1 

0  208 

0.250 

83 

17 

11.9 

u      j7 

10.2 

0.177 

0.208 

85 

15 

12.1 

"      18 

10.3 

0  384 

0  555 
(0.347) 

(13.5%) 

69 

31 

12.3 

Tyrosine  fed  as  on  May 
12. 

"      19 

10.1 

0.350 

0.815 
(0.607) 

(60.7%) 

43 

57 

12.5 

Phenol  fed  as  on  May 
10. 

cunivstances,  in  a  normal  dog.  This  is  in  accord  with  the  observa- 
tions of  Sweet  and  Ringer,^^  -^j^q  found  that  upon  giving  phlor- 
hizin  to  a  dog  with  Eck  fistula,  a  glycosuria  resulted  quite  com- 
parable to  that  occurring  in  a  normal  dog. 


Sweet,  J.  E.,  and  Ringer,  A.  I.,  ./.  Biol.  Chem.,  1913,  xiv,  1.35. 


Harry  Dubin 


79 


In  fasting,  as  was  to  be  expected,  there  was  a  diminution  in 
both  total  and  free  phenols,  the  latter  amounting  to  74  per  cent 
of  the  total.  A  greater  abiUty  to  conjugate  was  shown,  but  no 
initial  fall  and  subsequent  rise  in  the  phenol  excretion  as  described 
by  Herter*  and  Miiller^  was  noted.  Neither  was  it  possible,  by 
giving  calomel,  to  rid  the  urine  entirely  of  phenols.  In  this  con- 
nection, it  is  interesting  to  recall  that  Baumann,^^  owing  perhaps 
to  the  inadequacy  of  the  prevailing  methods,  could  get  no  test 
for  phenols  after  administering  calomel  to  a  fasting  dog. 

Experiment  G. — Dog  16-56  (Table  IV).  The  results  of  this 
experiment  are  in  accord  with  those  obtained  in  the  previous  Eck 
fistula  dog.  After  the  operation  there  was  a  tendency  for  the 
total  phenol  elimination  to  decrease,  due  perhaps  to  a  more  rapid 
rate  of  digestion.  The  influence  of  tyrosine  before  and  after  the 
operation  is  about  the  same — if  anything,  for  the  reason  just 
mentioned,  less  total  phenols  are  obtained  from  the  same  amount 
of  tyrosine  after  the  operation  than  before. 

Influence  of  Tyrosine,  Phenol,  and  p-Cresol  on  Phenol  Excretion  in 

Normal  Dogs. 

Experiments  K  and  M.— Dogs  15-74  and  16-20  (Table  V).  In 
general  the  findings  here  corroborated  those  of  Experiment  G. 


TABLE  V. 

Normal  Dog  15-74-. 

Influence  of  Tyrosine  on  Phenol  Excretion. 


Total 
N. 

Phenols. 

Weight. 

Date. 

i-d 

Remarks. 

Free. 

Total. 

P 

1915 

gm. 

g  m. 

gm. 

per 
cent 

per 
cent 

kg. 

Nov.  21 

12.1 

0.187 

0.217 

86 

14 

14.3 

"      22 

7.4 

0.187 

0.217 

86 

14 

14.4 

"      23 

12.1 

0.447 

0.610 
(0.393) 

(15.3%) 

73 

27 

14.4 

5.000  gm.  tyrosine  (2.570 
gm.  phenol)  given  in 
water  per  os. 

"      24 

12.8 

0.187 

0.217 

86 

14 

14.7 

"      25 

13.2 

0.189 

0.217 

87 

13 

14.8 

80  Physiology  of  Phenols 

TABLE  \— Concluded. 

Dog  16-20. 
Influence  of  Phenol  and  p-Cresol  on  Phenol  Excretion. 


Total 

N 

Phenols. 

Weight. 

Date. 

h-6 

Remarks. 

Free. 

Total. 

1916 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Feb.  20 

10.5 

0.158 

0.192 

83 

17 

11.1 

"      21 

6.3 

0.164 

0.195 

84 

16 

11.3 

"      22 

10.7 

0.167 

0.196 

85 

15 

11.1 

"      23 

10.5 

0.169 

0.196 

86 

14 

11.1 

"      24 

10.2 

0.166 

0.192 

86 

14 

11.1 

"      25 

9.9 

0.181 

0.647 
(0.455) 

(52.3%) 

41 

59 

11.2 

1.000  gm.  p-cresol  (0.870 
gm.  phenol)  given  in 
water  per  os. 

"      26 

10.1 

0.157 

0.192 

82 

18 

11.3 

"      27 

10.2 

0.417 

0.886 
(0.694) 

(69.4%) 

47 

53 

11.4 

1.000  gm.  phenol  given 
in  water  per  os. 

Exclusion  of  Pancreatic  Juice  from  the  Intestinal  Tract. 

Experiment  B-1. — Dog  15-63  (Table  VI).  The  normal  figures 
recorded  here  are  in  accord  with  those  of  Tables  II  and  V. 

Experiment  B-2. — Dog  15-63  (Table  VII).  This  experiment 
corroborates  fully  Experiment  A-3  (Table  III).  While  the  re- 
sults in  the  latter  experiment  were  observed  in  a  dog  with  Eck 
fistula,  those  of  the  present  experiment  were  noted  in  a  normal 
dog. 

Experiment  B-3. — Dog  15-63  (Table  VIII) .  Several  differences 
were  noted  between  the  results  of  this  experiment  and  those  of 
Experiment  B-2  (Table  VI).  After  the  operation,  the  total 
amount  of  phenols  eliminated  was  increased,  resulting  in  a  greater 
conjugation.  On  administering  tyrosine,  a  greater  percentage 
was  eliminated  as  phenols.  Undoubtedly  this  was  due  to  the 
fact  that  the  absence  of  pancreatic  juice  from  the  intestinal  tract 
retarded  the  processes  of  digestion.  The  administration  of  phe- 
nol and  of  7)-cresol  resulted  in  a  lessened  elimination;  i.e.,  with 
phenol,  there  was  a  drop  from  75.8  per  cent  to  51.1  per  cent  of 


Harry  Dubin 


81 


TABLE  VI. 
Exclusion  of  Pancreatic  Juice. 


Dog  Id- 


Influence  of  Phenol,   p-Cresol,   and  Tyrosine  on  Phenol  Excretion  before 

Operation. 


Total 

N. 

Phenols. 

Weight. 

Date. 

1  h-a 

Remarks. 

Free. 

Total. 

g1« 

6'^ 

191B 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Dec.    6 

12.7 

0.189 

0.223 

85 

15 

15.1 

7 

12.6 

0.192 

0.222 

86 

14 

15.1 

8 

13.1 

0.442 

0.980 

(0.758) 

(75.8%) 

45 

55 

15.2 

1.000  gm.  phenol  given 
in  water  per  os. 

9 

12.9 

0.195 

0.228 

86 

14 

"      10 

13.1 

0.195 

0.226 

86 

14 

15.3 

"    ■  11 

12.9 

0.411 

0.944 
(0.718) 

(71.8%) 

44 

56 

15.4 

Phenol  fed  as  on  Dec. 

8. 

"      12 

13,0 

0.195 

0.228 

86 

14 

15.2 

"      13 

13.9 

0.195 

0.224 

87 

13 

15.5 

"      14 

14.2 

0.215 

0.651 

(0.427) 

(49.2%) 

33 

67 

15.5 

1.000  gm.  p-cresol  (0.870 
gm.  phenol)  given  in 
water  per  os. 

"      15 

14.1 

0.195 

0.225 

87 

13 

15.5 

"      16 

14.2 

0.218 

0.606 
(0.381) 

(43.8%) 

36 

64 

15.4 

Cresol  fed  as  on  Dec.  14. 

u      17 

13.5 

0.195 

0.223 

87 

13 

15.6 

"      18 

14.5 

0.455 

0.625 
(0.402) 

(15.6%) 

73 

27 

15.6 

5.000  gm.  tyrosine  (2.57 
gm.  phenol)  given  in 
water  per  os. 

"      19 

13.3 

0.192 

0.222 

87 

13 

15.7 

THE  JOURNAL   OF   BIOLOGICAL   CHEMISTRY,    VOL.    XXVI,    NO.   1 


82 


Physiology  of  Phenols 


TABLE  VII. 
Exclusion  of  -Pancreatic  Juice.     Dog  15-63. 

Influence  of  Phlorhizin,  Fasting,  and  Catharsis  on  Phenol  Excretion  before 

Operation. 


Total 

N. 

Phenols. 

Weight. 

Date. 

i-o 

Remarks. 

Free. 

Total. 

0 

1915 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Dec.  19 

13.3 

0.192 

0.222 

87 

13 

15.7 

"      20 

14.8 

0.200 

0.268 

75 

25 

15.8 

1  gm.  phlorhizin. 

"      21 

15.5 

0.212 

0.278 

76 

24 

15.7 

1    «            ii 

"      22 

14.5 

0.192 

0.232 

83 

17 

15.6 

Sugar  in  urine  Dec.  20-23, 
inclusive. 

Animal  placed  on  kennel 
diet  till  Dec.  31,  when 
regular  diet  was  re- 
sumed. 

1916 

Jan.     5 

13.4 

0.188 

0.224 

84 

16 

16.0 

"        6 

4.6 

0.090 

0.130 

70 

30 

16.0 

Animal  fasting;  500  ce.  of 
water  given  in  cage. 

"        7 

3.6 

0.081 

0.107 

76 

24 

15.7 

"        8 

3.5 

0.080 

0.106 

76 

24 

15.4 

9 

3.2 

0.080 

0.106 

76 

24 

15.2 

"      10 

3.1 

0.079 

0.106 

75 

25 

14.9 

"      11 

3.3 

0.081 

0.107 

76 

24 

14.8 

"      12 

3  gm.  calomel  given;  cage 
urine  contaminated;  blad- 
der urine  contained  phe- 
nols. 

"      13 

3.1 

0  080 

0  106 

76 

24 

14.3 

"      14 

3.0 

0.081 

0.106 

76 

24 

14.1 

3  gm.  calomel  given. 

"      26 

11.1 

0  184 

0,223 

83 

17 

15.2 

Animal  on  regular  diet 
since  Jan.   15. 

Harry  Dubin 


83 


TABLE  VIII. 

Exclusion  of  Pancreatic  Juice.     Dog  15-63. 
Influence  of  Phenol,  p-Cresol,  and  Tyrosine  on  Phenol  Excretion  after 

Operation. 


Total 

N. 

Phenols. 

Weight. 

Date. 

1  13 

Remarks. 

Free. 

Total 

6 

u 

o 

1916 

gm. 

gm. 

gm. 

■per 
cent 

per 
cent 

kg. 

Jan.  26 

11.1 

0.184 

0.223 

83 

17 

15.2 

"      28 

Pancreatic  ducts  cut; 
dog  in  good  condi- 
tion. 

Feb.    6 

9.4 

0.200 

0.289 

69 

31 

15.5 

7 

10.0 

0.203 

0  293 

69 

31 

15.4 

"       8 

9.7 

0.202 

0.286 

71 

29 

15.3 

9 

9.9 

0.660 

0.845 
(0.559) 

(21.7%) 

78 

22 

15.4 

5.000  gm.  tyrosine  (2.57 
gm.  phenol)  mixed 
with  food. 

"      10 

9.5 

0.202 

0.289 

70 

30 

15.3 

"      11 

9.5 

0.200 

0.293 

68 

32 

15.5 

"      12 

10.1 

0.279 

0.632 
(0.339) 

(38.9%) 

44 

56 

15.5 

1.000  gm.  p-cresol  (0.870 
gm.  phenol)  given  in 
water  -per  os. 

"      13 

8.7 

0.203 

0.289 

70 

30 

15.3 

"      14 

8.5 

0.423 

0.800 
(0.511) 

(51.1%) 

53 

47 

15.3 

1.000  gm.  phenol  given 
in  water  per  os. 

"      15 

8.6 

0.200 

0.286 

70 

30 

15.2 

the  amount  fed;  with  p-cresol,  there  M^as  a  drop  from  49.2  per 
cent  to  38.9  per  cent  of  the  amount  given.  With  both  substances 
there  is,  however,  an  increased  conjugation  because  of  the  in- 
creased formation  of  phenols. 

Experiment  H. — Dog  16-55  (Table  IX).  The  results  of  this 
experiment  need  no  further  comment  since  they  corroborate  the 
previous  findings  as  recorded  in  Tables  VI  and  VIII. 


84 


Physiology  of  Phenols 


TABLE  IX. 


Exclusion  of  Pancreatic  Juice.     Dog  16-55. 

Influence  of  Phenol,  p-Cresol,  and  Tyrosine  on  Phenol  Excretion  before  and 

after  Section  of  Pancreatic  Ducts. 


e. 

Total 

N. 

Phenols. 

Weight. 

Dat 

JS? 

Remarks. 

Free. 

Total. 

6 

3   Qj 
O  bH 

1P16 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

May 

9 

9.5 

0.151 

0.200 

75 

25 

11.6 

(( 

10 

9.8 

0.417 

0.847 

(0.647) 

(64.7%) 

49 

51 

11.6 

1.000  gm.  phenol  given 
in  water  per  os. 

If 

11 

9.6 

0.156 

0.204 

76 

24 

11.7 

12 

9.6 

0.208 

0.594 
(0.390) 

(44.8%) 

35 

65 

11.8 

1.000  gm.  cresol  (0.870 
gm.  phenol)  given  in 
water  per  os. 

" 

13 

9.7 

0.153 

0.200 

76 

24 

11.8 

(( 

14 

9.8 

0.440 

0.625 
(0.425) 

(16.5%) 

70 

30 

11.8 

5.000  gm.  tyrosine  (2.570 
gm.  phenol)  mixed 
with  food. 

i< 

15 

Pancreatic  ducts  cut; 
dog  in  poor  condition. 

" 

16 

9.6 

0.175 

0.250 

70 

30 

11.3 

a 

17 

9.4 

0.173 

0.255 

68 

32 

11.0 

a 

19 

Animal  chloroformed. 

Intestinal  Obstruction. 

Experiment  C. — Dog  16-6  (Table  X).  The  results  obtained 
before  the  operation  are  in  accord  with  those  found  in  normal 
dogs.  After  the  operation,  it  is  worthy  of  note  that  although  the 
animal  ate  practically  nothing,  the  formation  and  elimination  of 
phenols  rose  to  a  high  level,  the  free  phenols  representing  only  56 
per  cent  of  the  total.  In  other  words,  the  conjugation,  due  to 
the  larger  amounts  of  phenols  present,  was  increased  just  as 
though  a  dose  of  phenol  had  been  ingested.  It  was  noted  also 
that  on  giving  phenol  after  the  operation,  only  51  per  cent  was 
eliminated,  while  before  the  operation  the  output  was  64  per 
cent.  This  is  explained  on  the  ground  that  digestion  is  markedly 
retarded.     In  this  respect  the  results  were  somewhat  similar  to 


Harry  Dubin 


85 


TABLE  X. 

Intestinal  Obstruction.     Dog  16-6.  " 

Influence  of  Phenol,  p-Cresol,  and  Tyrosine  on  Phenol  Excretion  before  and 
after  Intestinal  Obstruction. 


Total 

N. 

Phenols. 

Weight 

Date. 

j,"^ 

Remarks. 

Free. 

Total. 

6 
1 

0  w 

0 

1916 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Feb.  23 

9.6 

0.175 

0.227 

77 

23 

10.9 

"      24 

9.5 

0.171 

0.222 

77 

23 

11.1 

"      25 

9.7 

0.199 

0.548 
(0.326) 

(37.4%) 

36 

64 

11.1 

'  1.000  gm.  p-cresol  (0.870 
gm.  phenol)  given  in 
water  -per  as. 

"      26 

9.6 

0.173 

0.221 

78 

22 

11.2 

"      27 

9.5 

0.400 

0.862 

(0.641) 

(64.1%) 

46 

54 

11.1 

1.000  gm.  phenol  given 
in  water  per  os. 

"      28 

9.7 

0.174 

0.221 

79 

21 

11.3 

"      29 

9.8 

0.429 

0.605 

(0.384) 

(15.0%) 

71 

29 

11.4 

5.000  gm.  tyrosine  (2.57 
gm.  phenol)  mixed 
with  food. 

Mar.    1 

9.9 

0.173 

0.219 

79 

21 

11.3 

9 

Intestine  obstructed. 

"      10 

Urine  contaminated. 

"      11 

i(                       a 

"      23 

9.8 

0.263 

0.477 

55 

45 

Dog  defecated  hard 
stool  for  first  time 
since  operation. 

"      24 

7.2 

0.245 

0.442 

56 

44 

9.6 

Ate  only  part  of  diet. 

"      25 

6.4 

0.238 

0.403 

58 

42 

9.2 

No  food  given  Dec.  25 
and  Dec.  26. 

"      26 

4.9 

0.500 

0.914 
(0.511) 

(51.1%) 

55 

45 

8.9 

1.000  gm.  phenol  given 
in  water  per  os. 

those  obtained  before  and  after  the  exclusion  of  pancreatic  juice 
from  the  intestinal  tract.  On  March  27,  when  the  animal  was 
chloroformed,  the  intestine  was  found  to  be  dilated  to  about  100 
times  the  normal  capacity  for  about  3  feet  above  the  obstruction ; 
this  dilatation  continued  in  a  lesser  degree  up  to  the  jejunum. 
The  jejunum  and  duodenum  were  normal.  It  would  appear 
that  almost  complete  obstruction  had  been  obtained. 


86 


Physiology  of  Phenols 


Experiment  F. — Dog  15-63  (Table  XI).  The  findings  in  this 
experiment  were  not  as  marked  as  those  in  Experiment  C,  due 
perhaps  to  the  fact  that  the  amount  of  obstruction  secured  in 
this  case  was  not  as  great  as  in  Dog  16-6.  However,  there  was  a 
definite  rise  in  the  formation  and  elimination  of  phenols,  though 
nothing  unusual  was  to  be  noted  as  a  result  of  the  combination 
of  pancreatic  insufficiency  and  intestinal  obstruction.  Appar- 
ently the  increased  phenol  formation  was  not  sufficient  to  cause 
an  increase  in  the  conjugation,  the  free  phenols  representing 
almost  the  same  per  cent  of  the  total  both  before  and  after  the 
operation. 

TABLE  XI. 

Dog  15-63. 

Influence  of  Intestinal  Obstruction,  Combined  with  Pancreatic  Insufficiency, 
on  Phenol  Excretion. 


Total 

N. 

Phenols. 

Weight. 

Date. 

■  TJ 

Remarks. 

Free. 

Total. 

6 

d  is 
o  M 
O 

1916 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Apr.     8 

12.7 

0.190 

0.270 

70 

30 

13:9 

9 

12.9 

0.200 

0.263 

76 

24 

14.2 

"      10 

13.2 

0.498 

0.860 

58 

42 

14.2 

1.000  gm.  phenol  given 

"      11 

12.9 

0.357 

(0.597) 

(59.7%) 
0.588 

61 

39 

14.3 

in  water  per  os. 
1.000  gm.  p-cresol  (0.870 

"      12 

12.8 

0.187 

(0.325) 

(37.3%) 
0.257 

72 

28 

14.3 

gm.  phenol)  given  in 
water  per  os. 

"      13 

12.7 

0.195 

0.263 

74 

26 

14.3 

"      17 

Intestine  obstructed. 

"      19 

7.5 

0.227 

0.316 

71 

29 

13.7 

Dog  did  not  eat. 

"      20 

6.5 

0.543 

0.870 
(0.554) 

(55.4%) 

63 

37 

13.5 

"      "      "      "       1.000 
gm.   phenol  given  in 
water  per  os. 
No  lard  in  diet  begin- 

May   3 

10.9 

0.223 

0.307 

72 

28 

13.7 

ning  Apr.  22. 

"       4 

11.4 

0.555 

0.855 

65 

35 

13.8 

Phenol  fed  as  on  Apr. 

5 

11.3 

0.227 

(0.548) 

(54.8%) 
0.316 

72 

28 

13.7 

20. 

Harry  Dubin 


87 


Exclusion  of  Bile  from  the  Intestinal  Tract. 

Experiment  D. — Dog  16-39  (Table  XII).  The  figures  obtained 
before  the  operation  are  similar  to  those  in  a  normal  dog.  After 
the  operation,  however,  a  striking  difference  was  noted.     Although 


TABLE  XII. 

Exclusion  of  Bile.     Dog  16-39. 

Influence  of  Phenol,  p-Cresol,  and  Tyrosine  on  Phenol  Excretion  before  and 

after  Operation. 


e. 

Total 

N. 

Phenols. 

Weight. 

Dat 

1  -a 

Remarks. 

Free. 

Total. 

i 

§  M 

1916 

gm. 

gm. 

gm. 

per 
cent 

per 
cent 

kg. 

Mar. 

27 

13.6 

0.161 

0.215 

75 

25 

19.6 

« 

28 

13.8 

0.365 

0.758 
(0.543) 

(54.3%) 

48 

52 

19.9 

1.000  gm.  phenol  given 
in  water  per  os. 

i< 

29 

13.7 

0.167 

0.217 

76 

24 

20.3 

ii 

30 

13.9 

0.447 

0.613 
(0.396) 

(14.8%) 

72 

28 

20.5 

5.000  gm.  tyrosine  (2.570 
gm.  phenol)  mixed 
with  food. 

a 

31 

13.9 

0.189 

0.590 
(0.373) 

(42.8%) 

32 

68 

20.7 

1.000  gm.  p-cresol  (0.870 
gm.  phenol)  given  in 
water  per  os. 

Apr. 

1 

Bile  duct  cut;  dog  in 
good  condition. 

li 

2 

10.9 

0.167 

0  208 

80 

20 

20.5 

Bile  in  urine;  little  food 
eaten. 

ce 

3 

10.7 

0.330 

0.642 

(0.434) 
(43.4%) 

52 

48 

20.6 

Phenol  fed  as  on  Mar. 
28;  dog  ate  very  little; 
bile  in  urine. 

« 

7 

Dog  gave  birth. 

May 

4 

5.8 

0.238 

0.291 

82 

18 

11.8 

Animal  markedly  jaun- 
diced. 

11 

5 

6.4 

0.291 

0.513 
(0.222) 

(25.5%) 

57 

43 

11.5 

Cresol  fed  as  on  Mar.  31, 

u 

6 

6.5 

0.263 

0.329 

80 

20 

11.4 

u 

9 

6.7 

0.629 

0.839 
(0.510) 

(19.8%) 

75 

25 

Tyrosine  fed  as  on  Mar. 
30. 

88 


Physiology  of  Phenols 


there  was  an  increase  in  the  formation  and  elimination  of  phenols, 
the  free  phenols  represented  80  per  cent  of  the  total,  as  against  75  per 
cent  before  the  operation.  In  all  other  experiments,  an  increase  in 
the  phenol  formation  was  accompanied  hy  a  decrease  in  the  output 
of  free  phenols,  figured  as  per  cent  of  total,  while  in  this  experiment 
and  in  the  following  one  the  reverse  was  the  case.  Of  the  ingested 
phenol  and  p-cresol,  43.3  per  cent  of  the  former  and  25.5  per  cent 
of  the  latter  were  eliminated  after  the  operation  as  against  54.3 
and  42.8  per  cent,  respectively,  before.  Feeding  tyrosine  caused 
an  increased  conjugation,  the  free  phenols  dropping  from  75  to  72 
per  cent  before  the  operation,  and  from  80  to  75  per  cent  after. 
It  was  observed,  however,  that  only  14.8  per  cent  of  the  ingested 


TABLE  XIII. 


Exclusion  of  Bile.     Dog  16~41. 
Influence  of  Phenol,  p-Cresol,  and  Tyrosine  on  Phenol  Excretion  before  and 

after  Operation. 


Total 

N. 

Phenols. 

Weight. 

Date. 

1  TJ 

Remarks. 

Free. 

Total. 

6 

1916 

gm. 

gm. 

gm. 

per 

cent 

per 
cent 

kg. 

Apr.    8 

11.3 

0.151 

0.191 

79 

21 

9.6 

"        9 

11.1 

0.152 

0.186 

82 

18 

9.7 

"      10 

11.2 

0.477 

0.782 
(0.596) 

(59.6%) 

61 

39 

9.7 

1.000  gm.  phenol  given 
in  water  per  os. 

"      11 

11.2 

0.347 

0.568 
(0.382) 

(43.8%) 

61 

39 

9.7 

1.000  gm.  cresol  (0.870 
gm.  phenol)  given  in 
water  per  os. 

"       12 

11.1 

0.147 

0.182 

81 

19 

9.8 

"      1.3 

9.0 

0.440 

0.596 
(0.314) 

(12.2%) 

74 

26 

9.9 

5.000  gm.  tyrosine  (2.57 
gm.  phenol)  given  in 
water  per  os. 

"      14 

Bile  duct  cut;  animal 
in  poor  condition. 

"      15 

5,6 

0.098 

0.123 

80 

20 

8.9 

No  food  eaten;  bile  in 
urine. 

"      16 

5.2 

0.096 

0.120 

80 

20 

8.9 

No  food  eaten;  bile  in 
urine. 

"      17 

Animal  chloroformed. 

Harry  Dubin  89 

tyrosine  was  eliminated  before  the  operation  as  against  19.8  per 
cent  after.  This  was  similar  to  the  result  obtained  with  tyrosine 
after  excluding  the  pancreatic  juice,  and,  as  in  the  latter  case, 
may  be  explained  on  the  basis  of  delayed  digestion. 

Experiment  E.—Bog  16-41  (Table  XIII).  The  results  of  this 
experiment  were  not  as  clean  cut  as  they  might  be,  for  the  animal 
refused  to  eat  and  was  rather  sick.  On  autopsy  the  bile  duct, 
up  to  the  point  tied,  was  greatly  enlarged. 

DISCUSSION. 

The  formation  of  phenolsulf uric  acids  is  one  example  of  the 
abihty  of  the  body  to  convert  poisonous  substances  into  harmless 
compounds.  It  is,  however,  an  open  question  as  to  whether  this 
power  of  the  body  is  sufficient  for  all  pm-poses.  Since  the  time  of 
Baumann  it  has  been  believed  that  phenols  were  quantitatively 
converted  into  harmless  phenol  esters;  hence  phenols  were  esti- 
mated on  the  basis  of  conjugated  sulfates,  whereas  our  figures 
corroborate  the  statement  made  by  Folin^  that  the  greater  part  of 
the  phenols  are  excreted  unconjugated. 

It  has  been  thought  that  the  free  phenols  are  harmful  to  the 
body,  and  this  must  be  due  primarily  to  an  increase  of  total 
phenols,  with  its  accompanying  increase  of  free  phenols.  For 
example,  it  is  seen  in  normal  cases  that  the  absolute  amount  of 
free  phenols  represents  from  75  to  85  per  cent  of  the  total.  On  the 
other  hand,  where  there  is  a  rise  in  total  phenols,  whether  due  to 
ingested  phenols  or  to  some  pathological  condition,  the  free  phe- 
nols, although  increased  absolutely,  represent  from  30  to  70  per 
cent  of  the  total.  In  other  words,  the  absolute  amount  of  free 
phenols  should  be  taken  into  consideration  together  with  the  per- 
centage in  considering  their  deleterious  infiuence. 

With  one  exception,  an  increase  in  the  formation  of  phenols, 
whether  normal  or  pathological,  resulted  in  an  increased  conjuga- 
tion, though  the  reverse  might  have  been  expected.  It  would 
appear  that  the  protective  mechanism  of  the  body  responded  to 
the  greater  stimulus.  The  exception  noted  was  in  the  case  where 
bile  was  excluded  from  the  intestinal  tract.  Here  there  was  an 
increased  phenol  formation  accompanied  by  a  decreased  conju- 
gation. It  would  seem,  therefore,  that  the  bile  plays  some  part 
in  assisting  the  liver  in  its  conjugating  function. 


90  Physiology  of  Phenols 

There  is  a  greater  conjugation  after  feeding  p-cresol  than  after 
giving  phenol.  In  view  of  the  fact  that  p-cresol  makes  up  the 
larger  part  of  the  urinary  phenols,  it  seems  plausible  to  assimie 
that  when  it  is  introduced  into  the  body  it  is  more  easily  conju- 
gated. Hence  on  feeding  this  substance  the  elimination  of  free 
phenols  rises  only  very  little. 

The  high  percentage  of  free  phenols  prevailing  after  Eck  fistula 
was  to  be  expected,  inasmuch  as  the  liver,  which  is  the  main  seat 
of  conjugation,  has  been  cut  out  of  the  circulation.  In  spite  of 
this,  upon  the  ingestion  of  phenol  there  is  an  increased  conjuga- 
tion, showing  that  other  organs,  when  necessary,  can  take  up  the 
work  of  the  liver  in  this  respect. 

Lewin^  held  that  the  increased  protein  breakdown  accounted 
for  the  increased  phenol  formation  when  phlorhizin  was  given. 
While  it  is  true  that  phlorhizin  causes  a  protein  breakdown,  it  is 
highly  improbable  that  this  is  the  reason  for  an  increased  phenol 
production.  Regarding  the  fate  of  phlorhizin  in  the  body,  it  has 
been  shown^^  that  when  injected,  part  of  it  is  eliminated  as  a 
combined  glucoronic  acid  while  another  part  apparently  under- 
goes further  change;  also,  unchanged  phlorhizin  can  be  found  for 
some  time  in  the  blood  and  tissues.  In  view  of  all  this,  when  it 
is  considered  that  phlorhizin  contains  two  benzene  radicals,  and 
that  benzene  when  fed^  gives  rise  to  phenol,  it  seems  likely  that  it 
is  the  phlorhizin  per  se  which  is  converted  into  phenol. 

The  results  obtained  show,  among  other  things,  that  gastro- 
intestinal disturbances,  in  which  it  is  reasonable  to  assume  that 
there  is  an  increase  in  the  formation  of  phenol,  and  also  certain 
liver  diseases,  where  it  is  supposed  that  the  mechanism  for  the 
detoxication  of  phenols  has  been  impaired,  offer  a  field  for  inves- 
tigation, for,  as  Folin^  points  out,  and  as  this  work  proves,  the 
phenols  can  be  taken  as  an  index  of  intestinal  putrefaction. 

SUMMARY.    , 

The  results  of  this  investigation,  in  addition  to  confirming  the 
findings  of  Folin,^  bring  to  light  some  interesting  observations. 

A.  1.  The  elimination  of  phenols  from  day  to  day  is  quite 
constant. 

«  VonFiirth,  O.,  Chemistry  of  Metabolism,  Philadelphia,  1916,  280. 


Harry  Dubin  91 

2.  Withdrawing  water  from  the  diet  causes  an  increased  phenol 
elimination. 

3.  After  Eck  fistula,  the  free  phenols  represent  as  high  as  97 
per  cent  of  the  total,  the  latter,  as  also  the  former,  exhibiting  a 
tendency  to  decrease. 

4.  After  intestinal  obstruction,  there  is  an  increase  in  both  free 
and  total  phenols,  the  former  constituting  as  low  as  55  per  cent  of 
the  latter. 

5.  In  pancreatic  insufficiency,  there  is  an  increase  in  both  free 
and  total  phenols  with  an  accompanying  decrease  in  the  percent- 
age of  free  phenols. 

6.  After  excluding  the  bile,  both  free  and  total  phenols  increase, 
but  with  an  increase  in  the  percentage  of  free  phenols. 

B.  1.  The  feeding  of  phenol  and  2?-cresol  results  normally,  with 
but  slight  variations,  in  the  elimination  of  about  G5  per  cent  and 
40  per  cent  respectively. 

2.  After  Eck  fistula,  about  the  same  excretion  is  noted. 

3.  After  intestinal  obstruction,  pancreatic  insufficiency,  and  ex- 
clusion of  bile,  there  is  in  both  cases  a  drop  in  the  amount  excreted. 

C.  1.  The  feeding  of  tyrosine  results  normally  in  an  excretion 
of  about  14  per  cent,  as  phenols. 

2.  After  Eck  fistula,  practically  the  same  amount  is  eliminated. 

3.  After  exclusion  of  bile  and  pancreatic  juice,  about  20  per 
cent  of  ingested  tyrosine  is  eliminated. 

D.  Feeding  of  any  of  the  three  substances  causes  an  increase  in 
the  conjugation  at  all  times — p-cresol  to  a  greater  degree  than 
phenol,  and  the  latter  to  a  greater  extent  than  tyrosine. 

E.  Tyrosine,  phenol,  and  p-cresol,  fed  in  amounts  of  5  gm.,  1 
gm.,  and  1  gm*.  respectively,  were  all  eliminated  within  24  hours. 

F.  No  unchanged  tyrosine  could  be  demonstrated  in  the  urine 
or  feces. 

G.  Fasting  reduces  the  phenols  to  a  low  level;  the  injection  of 
phlorhizin  during  fasting  causes  an  increase  in  the  output  of 
phenols. 

H.  It  is  impossible  to  free  the  urine  entirely  of  phenols  by  the 
use  of  calomel. 

I.  The  bile  appears  to  have  some  influence  on  the  conjugating 
function  of  the  liver. 

J.  The  phenols  can  be  taken  as  an  index  of  intestinal  putre- 
faction. 


THE  WAVERLY  PRESS 
BALTIMORK.  U.  S.  A. 


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